PROJECT SUMMARY/ABSTRACT This proposal requests funding for the National Center for Microscopy and Imaging Research (NCMIR) at UCSD to acquire a 300-keV FEI state-of-the-art intermediate-high voltage electron microscope (IVEM) and integrate it with massive data storage and computational resources. The result, to benefit the research and technology development aims of NIH biomedical investigators, will be an integrated big data appliance for volume visualization (to nanometer resolution) through extensive three-dimensional (3D) volumes. This coupled microscope / computational system will be equipped to perform automated large-volume 3D imaging required for a range of biomedical research projects, including imaging of whole cells at resolutions sufficient to pinpoint the molecular building blocks comprising macromolecular complexes, intra-nuclear and cytoplasmic structures, and characterization of nanostructures of biological materials in the contexts of their functional environments. This proposal includes eight major and three minor research projects that exemplify the transformative science that can be accomplished with this new one-of-a-kind instrument. Because this instrument will generate massive 3D datasets requiring substantial resources for computation-based refinement and quantitative analysis, it will be coupled tightly to a large, scalable data- storage and high-performance computing environment, leveraging the significant resources at the San Diego Supercomputer Center and the California Institute for Telecommunications and Information Technology (both at UCSD). Prominent national, international, and local users will broaden the application of this platform to achieve maximum impact in advancing scientific discovery and human health. They will work together and with our management team to commission, use, and refine the capabilities of this microscope. They will also help others benefit fully from its capabilities. Our experiences in designing and commissioning high-end instruments and those of our staff and scientists using them have been and will be communicated to the manufacturers with the goal of guiding the advance of the capabilities of future instruments in directions of most benefit to the biomedical research community.